Relationship between quantitative real-time PCR cycle threshold and culture for detection of subspecies .

Can Vet J

Department of Pathobiology and Centre for Public Health and Zoonoses, Ontario Veterinary College, University of Guelph, 50 Stone Road East, Guelph, Ontario NIG 2W1 (Weese); Diagnostic Services (Saab) and Department of Health Management (Saab, McClure), Atlantic Veterinary College, University of Prince Edward Island, 550 University Avenue, Charlottetown, Prince Edward Island C1A 4P3; Animal Health and Welfare Branch, Ontario Ministry of Agriculture, Food and Rural Affairs, 5th Floor NW, 1 Stone Road West, Guelph, Ontario N1G 4Y2 (Moore); Animal Health Laboratory, University of Guelph, 419 Gordon Street, Guelph, Ontario N1G 2W1 (Cai).

Published: June 2023

Objective: To compare PCR and culture results for the detection of subspecies .

Animals: Respiratory tract samples (N = 158) from horses being tested for

Procedure: Bacterial culture was carried out on samples from which was detected by quantitative real-time PCR.

Results: was isolated from 12 (7.6%) samples: 4/9 (44%) samples when the PCR cycle threshold (C) was ≤ 30, 7/30 (23%) when the C was 30.1 to 35, and 1/119 (0.8%) when the C was 35.1 to 40. The highest C sample from a sample that yielded a positive culture was 36.9. The optimal Youden's J value was at a C of 34.2, the same value as determined by number needed to misdiagnose when the cost of a false negative is deemed to be either 5 or 10 × that of a false positive.

Conclusions: Viable was only detected in a minority of quantitative PCR (qPCR) positive samples. A qPCR C of 34.2 was a reasonable breakpoint for likelihood of the presence of culturable

Clinical Relevance: Evaluation of C values may be useful as a proxy to indicate the likelihood of cultivable being present and could be useful as part of risk assessments.

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Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC10204887PMC

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